Solution structure of Blo t 19, a minor dust mite allergen from Blomia tropicalis
The data from this entry were used to determine structure of Blo t 19. Due to Proline isomerization, Blo t 19 spectrum has degenerate resonances for a stretch of six residues in its C-terminus. These two populations have varying intensities and are unambiguously assigned as major and minor population. Solution structure is calculated for the Major population.
FIRST TWO RESIDUES (GS) ARE EXPRESSION TAGS.
NMR data was acquired at 295K using Shigemi NMR tubes.
Initial structure ensemble was calculated by semi-automated NOESY assignment by CYANA. The assignments were manually verified in Sparky and final structure annealing was performed in CYANA. Structure and restraints from CYANA were imported in Xplor-NIH for explicit water refinement.
torsion angle dynamics, DGSA-distance geometry simulated annealing
1 mM [U-100% 13C; U-100% 15N] Blo t 19-1, 50 mM potassium phosphate-2, 100 mM sodium chloride-3, 90% H2O/10% D2O
1 mM [U-100% 15N] Blo t 19-4, 50 mM potassium phosphate-5, 100 mM sodium chloride-6, 90% H2O/10% D2O
1 mM Blo t 19-7, 50 mM potassium phosphate-8, 100 mM sodium chloride-9, 90% H2O/10% D2O
1 mM [U-100% 15N] Blo t 19-10, 50 mM potassium phosphate-11, 100 mM sodium chloride-12, 15 mg/ml Pf1 phage-13, 90% H2O/10% D2O
Schwieters, Kuszewski, Tjandra and Clore